title: Methylation of an intronic region regulates miR-199a in testicular tumor malignancy.
abstract: In the testicular cancer cell line, NT2, we previously demonstrated that differentially methylated regions were located in introns or intergenic regions, and postulated these might regulate non-coding RNAs. Three microRNAs and three small nucleolar RNAs were differentially methylated; one, miR-199a, was associated with the progression and prognosis of gastric and ovarian cancers. In this report we document, by epigenomic profiling of testicular tissue, that miR-199a is transcribed as antisense of dynamin 3 (chromosome 1q24.3), and hypermethylation of this region is correlated with miR-199a-5p/3p repression and tumor malignancy. Re-expression of miR-199a in testicular cancer cells led to suppression of cell growth, cancer migration, invasion and metastasis. The miR-199a-5p, one of two mature miRNA species derived from miR-199a, is associated with tumor malignancy. We further identified the embryonal carcinoma antigen podocalyxin-like protein 1 (PODXL), an anti-adhesive protein expressed in aggressive tumors, as a target of miR-199a-5p. We demonstrated PODXL is overexpressed in malignant testicular tumor, and cellular depletion of PODXL resulted in suppression of cancer invasion. The inverse relationship between PODXL and miR-199a-5p expression suggests PODXL is a downstream effector mediating the action of miR199a-5p. This report identifies DNA methylation, miR-199a dysregulation and PODXL as critical factors in tumor malignancy.
authors.affiliation: Section on Clinical and Developmental Genomics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
authors.forename: H-H
authors.initials: HH
authors.lastname: Cheung
authors.forename: A J
authors.initials: AJ
authors.lastname: Davis
authors.forename: T-L
authors.initials: TL
authors.lastname: Lee
authors.forename: A L
authors.initials: AL
authors.lastname: Pang
authors.forename: S
authors.initials: S
authors.lastname: Nagrani
authors.forename: O M
authors.initials: OM
authors.lastname: Rennert
authors.forename: W-Y
authors.initials: WY
authors.lastname: Chan
mesh_terms: D000818:Animals; D002277:Carcinoma / Q000235:genetics* / Q000556:secondary; D045744:Cell Line, Tumor; D002465:Cell Movement; D019175:DNA Methylation; D034286:Dynamin III / Q000096:biosynthesis; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007438:Introns; D008113:Liver Neoplasms / Q000556:secondary; D008175:Lung Neoplasms / Q000556:secondary; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis*; D009361:Neoplasm Invasiveness; D012795:Sialoglycoproteins / Q000235:genetics; D013736:Testicular Neoplasms / Q000235:genetics* / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: D035683:MicroRNAs; D012795:Sialoglycoproteins; C531408:mirn199 microRNA, human; C001643:podocalyxin; D034286:Dynamin III